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Epidermodysplasia verruciformis human papillomavirus (EV-HPV) DNA has been demonstrated in malignant and benign skin lesions and in hairs plucked from renal transplant recipients and immunocompetent patients. We investigated the association between EV-HPV DNA in hairs plucked from eyebrows and the occurrence of non-melanoma skin cancer (NMSC) in a community-based study. Within a cohort of residents of a Queensland township (Nambour), nested case-control studies of recently developed NMSC (64 cases), basal-cell carcinoma (BCC) (51 cases) and squamous-cell carcinoma (SCC) (25 cases) were conducted. EV-HPV DNA in hair and a small number of available tumour samples was detected using a nested PCR specific for EV-HPV types. EV- HPV DNA was detected in hairs from 94 of 143 individuals (66%), and 36 (39%) of the samples contained 2 or more different EV-HPV types. Only known or putatively new EV-HPV types were detectable after sequencing 93 samples. EV- HPV status agreed for 12 of 20 subjects who had both hair and skin tumour samples available. In 4 of 5 pairs of positive samples, the same EV-HPV type was found. There were non-significant negative associations between EV-HPV and NMSC (OR 0.77, 95% CI 0.34-1.8) and BCC (OR 0.58, 95% CI 0.23-1.5) but a non-significant positive association with SCC (OR 2.00, 95% CI 0.50-8.0). (C) 2000 Wiley-Liss, Inc.